Differential effects of actinomycin d and cordycepin in lettuce seed germination and RNA synthesis

Intact lettuce seed germination was inhibited by cordycepin but not by actinomycin D; however, when seeds were clipped at the cotyledonary end, actinomycin D partially inhibited germination. Uptake studies with intact seeds using ³H-actinomycin D showed that it was unable to reach the embryo prior to radical protrusion. ³H-Cordycepin uptake studies using intact seeds showed that cordycepin was able to reach the embryo during the first 3 hours of incubation and at subsequent times. The pericarp and endosperm offered resistance to penetration of cordycepin into the embryo. In contrast to actinomycin D, cordycepin markedly inhibited ³H-uridine incorporation into RNA of intact seeds during the first 10 and 12 hours of incubation. About 60% of ³H-adenosine incorporation into poly A-RNA was inhibited by cordycepin during 12 hours of incubation, whereas actinomycin D had little effect. RNA synthesis appears to be essential for seed germination.     

Einfluss von hydroxyharnstoff auf die DNA-synthese und keimung der samen vonAgrostemma githago sowieVaccaria pyramidata

Application of hydroxyurea to intact seeds ofAgrostemma githago as well asVaccaria pyramidata causes a strong inhibition of radicular³H-thymidine incorporation into DNA, but germination rate is only insignificantly reduced. The seedling development, however, is strongly affected. Consequently, DNA synthesis is not a prerequisite for the radicle protrusion.     

STUDIES ON THE GERMINATION OF SEEDS OF THE ROOT PARASITES,ALECTRA VOGELII AND STRIGA GESNERIOIDES. II. DNA SYNTHESIS AND DEVELOPMENT OF THE QUIESCENT CENTER IN THE RADICLE

DNA synthetic activity in the radicle meristem of embryos of germinating seeds of the obligate root parasites, Alectra vogelii and Striga gesnerioides was followed by autoradiography of ³H-thymidine incorporation. Incorporation of ³H-thymidine occurred in the nuclei of cells destined to form the vascular tissues, ground meristem and epidermis. An analysis of the distribution of labeled nuclei demonstrated the presence of a quiescent center of 2-4 cells in the radicle at the beginning of seed germination, becoming more prominent at later stages of germination. During continued growth of the radicle which resulted in a reduction in size of the meristem, cells of the original quiescent center were activated to undergo DNA synthesis.     

Release of Esterase Following Germination of Lettuce Seed (Lactuca sativa L.)

Light-insensitive lettuce seeds, Lactuca sativa L. cv. Great Lakes, release esterases for a period following radicle protrusion. Very little or no enzymes are released prior to 24 hours or after 48 hours of germination. As compared to intact seeds, half-seeds readily release esterases and the release is not affected by far red irradiation. Bulk of the released esterases are derived from the endosperm tissue and presumably exists in the intact seed as a component of the extraembryonic fluid.     

Dormancy in Dioscorea: Rapid Germination of Detached Embryos from Dormant Seeds of D. tokoro

The intact dormant seeds of Dioscorea tokoro germinate slowlyif at all between 11-23°C; for full and rapid germinationthey require prior chilling treatment [Okagami and Kawai (1982)Bot. Mag. Tokyo 95: 155]. The germination abilities of zygoticembryos detached from dormant seeds of this species were studiedunder various nutritional and temperature regimes. For germinationof embryos, the minimum nutritional components in Murashigeand Skoog's (1962) medium that were required were sucrose andNO^–₃ or SO^2–₄. As the source of carbohydrate forgermination of detached embryos, sucrose, mannose and maltosewere effective; glucose and fructose were less effective; andrhamnose was entirely unable to support germination. Embryos detached from dormant seeds, incubated with the sucroseplus KNO₃, germinated more rapidly with increasing temperatureup to 35°C. However, application of sucrose and KNO₃ didnot induce germination of intact seeds above 26°C. Therefore,it is very possible that the endosperm exerts an inhibitoryfunction on germination at such high temperatures. When seeds were incubated after a cut was made over a smallpart of the edge of the endosperm in which the radicle of theembryo is encased, germination occurred rapidly but the increasein germination percentage was slight. This result suggests thatthe endosperm suppots part of the germination inhibition bymeans of a mechanical barrier or its impermeability to wateror gases. Physiological features of the endosperm alone or interactionsbetween the embryo and endosperm may contribute significantlyto the characteristics of dormancy of intact seeds of this species. (Received May 30, 1988; Accepted January 11, 1989)     

Activation of the Cell Cycle in Tomato (Lycopersicon esculentum Mill.) Seeds during Osmoconditioning as Related to Temperature and Oxygen

Using flow cytometric analyses of the nuclear DNA content, westudied the effects of various conditions of osmopriming onthe activation of the cell cycle in embryo root tips of tomato(Lycopersicon esculentum‘Elko’) seeds. In dry untreatedseeds, 90.7% of the nuclei revealed 2C signals. Priming of seedsin polyethylene glycol-8000 (PEG) improved the germination rateof seeds transferred onto water at 15 °C. This was associatedwith an increase in 4C signals when priming was carried outat -1.0 and -1.5 MPa. Priming at -2.0 MPa enhanced subsequentgermination but had no effect on DNA replication. For temperaturesduring priming up to 25 °C, a positive linear correlationexisted between the efficiency of the treatment, evaluated bythe reciprocal of time to obtain 50% germination at 15 °C,and the frequency of 4C nuclei or the 4C/2C values. Such a correlationdid not exist when priming was performed at higher temperatures.At least 5% oxygen in the atmosphere was required during primingfor the induction of DNA replication and for the enhancementof subsequent germination. In the presence of 5x10^-4M and 10^-3MNaN₃during priming, most of the cells were maintained with 2CDNA levels and the treatment had no stimulatory effect on germination.The results show a positive linear relationship between thefrequency of 4C DNA nuclei or the 4C/2C ratio and the improvingeffect of priming. However, in suboptimal conditions of priming(-2.0 MPa or temperatures higher than 25 °C), the improvementof seed germination was not associated with the onset of DNAreplication.Copyright 1999 Annals of Botany Company Cell cycle, germination, osmopriming, oxygen, temperature, Lycopersicon esculentum, tomato.     

Primary dormancy in tomato (Lycopersicon esculentum cv. Moneymaker): studies with the sitiens mutant

Intact wild-type tomato (Lycopersicon esculentum cv. Moneymaker)seeds do not complete germination to the same percentage orat the same speed as intact ABA-deficient sitiens (sit^w) mutantseeds when seeds of both genotypes are imbibed on polyethyleneglycol (PEG) solutions of –0.3 to –1.5 MPa osmoticpotential. However, if the thicker testas of wild-type seedsare removed (stripped) from the micropyle without damaging theendosperm, both the percentage and speed of germination at lowexternal water potential are similar to that of sit^w mutantseeds. Removing the micropylar end of the testa from sit^w seedsdid not enhance either the speed or percentage of germinationon PEG solution. Despite similar germination percentage and speed between strippedwild-type seeds and either stripped or intact sit^w seeds underosmotic stress, some differences in seed metabolism are evidentbetween genotypes. The activity of endo-ß-mannanasewas greater in the endosperm of sit^w mutant seeds compared tothe endosperm of wild-type seeds when seeds were exposed toosmotic stress. Although     

鹭兰种子无菌繁殖球根

为建立鹭兰（Habenaria radiata）种子繁殖法,对无菌培养条件下的种子发芽和幼苗生长进行了观测,并对无菌繁殖获得的球根进行了种植试验。结果表明,鹭兰种子不进行处理难以无菌培养发芽,种子通过75%乙醇10 s+1% NaClO 10 min处理,或者40 kHz超声波2.5 min+1% NaClO 10 min处理,在1/2MS培养基的发芽率可达58%以上;在1/2MS培养基中,添加GA-3 1 mg·L^-1对叶片生长有促进作用,添加IAA 1 mg·L^-1对根生长和球根形成有促进作用,球根形成率达55%;无菌繁殖的球根贮存至春季盆栽,成苗率约72%。     

Investigating seed dormancy in cotton (Gossypium hirsutum L.): understanding the physiological changes in embryo during after‐ripening and germination

• The dormancy of seeds of upland cotton can be broken during dry after‐ripening, but the mechanism of its dormancy release remains unclear.
• Freshly harvested cotton seeds were subjected to after‐ripening for 180 days. Cotton seeds from different days of after‐ripening (DAR) were sampled for dynamic physiological determination and germination tests. The intact seeds and isolated embryos were germinated to assess effects of the seed coat on embryo germination. Content of H₂O₂ and phytohormones and activities of antioxidant enzymes and glucose‐6‐phosphate dehydrogenase were measured during after‐ripening and germination.
• Germination of intact seeds increased from 7% upon harvest to 96% at 30 DAR, while embryo germination improved from an initial rate of 82% to 100% after 14 DAR. Based on T₅₀ (time when 50% of seeds germinate) and germination index, the intact seed and isolated embryo needed 30 and 21 DAR, respectively, to acquire relatively stable germination. The content of H₂O₂ increased during after‐ripening and continued to increase within the first few hours of imbibition, along with a decrease in abscisic acid (ABA) content. A noticeable increase was observed in gibberellic acid content during germination when ABA content decreased to a lower level. Coat removal treatment accelerated embryo absorption of water, which further improved the accumulation of H₂O₂ and changed peroxidase content during germination.
• For cotton seed, the alleviation of coat‐imposed dormancy required 30 days of after‐ripening, accompanied by rapid dormancy release (within 21 DAR) in naked embryos. H₂O₂ acted as a core link between the response to environmental changes and induction of other physiological changes for breaking seed dormancy.

     

Total Uptake and Incorporation into DNA of 3H-Thymidine, 3H-Deoxyuridine, and 3H-Thymine in the Primary Root of Vicia faba L.: II. EXCISED ROOTS

Total uptake of ³H-thymidine, ³H-thymine, and ³H-deoxyuridineand incorporation of these substances into DNA have been investigatedin excised roots in Vicia faba. Total uptake was found to behigher in excised roots than in intact ones. This was a consequenceof exogenous ³H-DNA precursors entering the excised roots throughthe cut surface. Where the cut surface was not immersed in the³H-thymidine solution ³H uptake was also higher than in intactroots. In this case ³H uptake scemed to be correlated with water-lossfrom the cut surface. Incorporation of ³H-thymidine, ³H-thymine,and ³H-deoxyuridine into DNA was found to be higher in excisedroots than in intact ones, probably as a result of a decreasein the size of the relevant endogenous precursor pools. It issuggested that this decrease resulted from the cessation ofthe supply of DNA precursors to these roots from the cotyledons.     

防风种子发芽特性及促进发芽的试验研究

防风种子发芽率和发芽势均较低,其主要原因包括个体之间的差异及物种特性。种子发芽率较低首先表现在个体发芽率上的差异,试验所用15株防风发芽率从28.0%到92.0%,从整体 上表现发芽率较低。防风本身发芽率低,由于物种因素,种子刚刚采收后的休眠,个体之间解除休眠时间和进入衰老的时间不一致,致使所有的种子不能全部在同一时间进入发芽高峰,同时,防风发芽的启动日有所差异,15株防风启动日和发芽持续时间相差均为两周以上。采用5~50 mg·kg^-1 GA、1~10 mg·kg^-1 6-BA、1%KNO₃和3%H₂O₂可提高休眠防风种子的发芽率,GA、6-BA可除防风种子的休眠,而1% KNO₃和3% H₂O₂对解除休眠的种子无明显影响。采用多种微量元素,即10~100 mg·kg^-1 Mn²⁺、10 mg·kg^-1 Cu²⁺和1 mg·kg^-1 Mo对种子进行处理可显著提高防风种子的发芽率,提示在植株绿果期喷施该类元素也可提高防风种子的发芽率。采用GA、NAA处理幼果,也可以提高种子的发芽率。     

Synergism between gibberellic acid and low Pfr levels inducing germination of Kalanchoë seeds

Sown on water, seeds of Kalanchoëbiossfetdiana Poelln. cv. Feuerblute are absolutely light-requiring and show full red/far-red reversibility. In seeds, sown on 2 ×10^-3 M gibberellic acid, red/far-red reversibility disappears and both short red and far-red irradiations induce germination. Gibberellic acid alone does not induce germination, but it increases the physiological activity of P_fr to the extent, that the low P_fr level obtained by far-red irradiation becomes very effective. The synergism between gibberellic acid and far-red light appears after a two-day incubation; period. The nature of this lag phase was examined by measuring both germination and uptake of labelled gibberellic acid in intact seeds and seeds with a punctured seed coat. The lag phase was shown to be independent of the uptake kinetics of gibberellic acid and allows development to a specific stage, necessary for germination after phytochrome-phototransformation. The kinetics of the uptake of gibberellic acid by intact seeds and embryos of intact seeds are different. In intact seeds most of the gibberellic acid is retained in the seed coat; only a small fraction actually penetrates to the embryo where it can exert its physiological activity.     

不同光照处理下青藏高原克隆植物黄帚橐吾(Ligulariavirgaurea)种子大小对其幼苗生长的影响

以高寒草甸克隆植物黄帚橐吾为实验材料,通过遮荫网模拟植被遮荫,研究种子大小与萌发及幼苗生长能力的关系和幼苗对光照条件的反应。结果表明：（1）在自然光照下,黄帚橐吾种子大小对种子萌发的影响显著,大种子的萌发率高于小种子。遮荫生境下,大、小种子萌发率有所降低,但遮荫对小种子萌发的影响比大种子显著。小种子的萌发率下降了近1/8,而大种子的萌发率仅下降了1/11。（2）黄帚橐吾种子大小对幼苗生物量积累影响显著,大种子幼苗总生物量（TB）大于小种子幼苗的。但生物量的分配与播种时间相关,播种后60 d,在自然光照条件下,大种子幼苗对根生物量的分配大于小种子幼苗,而对叶生物量的分配则正好相反。在遮荫环境中,大、小种子幼苗普遍对根的生物量分配增加,大种子幼苗根冠比（R/S）大于小种子幼苗。（3）黄帚橐吾种子大小对幼苗的生长也有明显影响。在自然光照下,小种子幼苗的相对生长速率（RGR）较大于大种子幼苗,但叶面积比率（LAR）、叶面积干质量比（SLA）、叶干质量（LWR）差别不明显。在遮荫条件下,幼苗的LAR、SLA、LWR显著增加,但大、小种子幼苗间差异不显著,幼苗的RGR减小,小种子幼苗的减小趋势大于大种子幼苗。     

Amounts of Nuclear DNA and Internal Morphology of Gibberellin- and Abscisic Acid-deficient Tomato (Lycopersicon esculentum Mill.) Seeds During Maturation, Imbibition and Germination

Using X-ray photography and flow cytometry, the internal morphologyand DNA replication activity of wild type (wt), GA- (gib-1 )and ABA-deficient (sit^w ) tomato (Lycopersicon esculentum Mill.cv. Moneymaker) mutant seeds were studied. During seed formation,from 30 to 45 d after pollination (DAP) the endosperm becomessolid and the seed starts to gain desiccation tolerance. Atthis time significant changes occur in the amounts of DNA inradicle tip cells. At 30 DAP, radicle tip cells of the threegenotypes manifest about 60% of 2C, 30% of 4C and 10% of 8Camounts of DNA. Upon maturation (45 DAP onwards), most cellsin the seeds of the three genotypes arrest in the G₁phase ofthe cell-cycle with 2C amounts of DNA. However, a relativelyhigh proportion of cells with 4C amounts of DNA was detectedin the radicle tip cells ofsit^w compared with wild type andgib-1. At the well-matured stage (60 DAP), there were about 2% ofseeds with free space in wild type andgib-1 , and about 13%insit^w . At the over-matured stage (75 DAP), even more seedswith free space were found insit^w , whereas no increase in theproportion of the seeds with free space was detected in theother two genotypes. In -1.0 MPa PEG-6000 with or without 10µM GA₄₊₇, no germination occurred in well-matured wildtype andgib-1 seeds, whether or not they were dried after harvest.However,sit^w seeds were able to germinate both in over-maturefruit and in -1.0 MPa PEG-6000. Priming of dried seeds in -1.0MPa PEG induced a large amount of free space in almost all seedsof the three genotypes, and nuclear DNA synthesis in the radicletip cells of wild type andsit^w seeds. However, PEG priming offresh (non-dried) seeds had no effect on the amount of freespace and 2C/4C DNA ratios in wild type orgib-1 seeds, but didinduce free space in about 20–25% ofsit^w seeds and provoked4C signals insit^w seeds. Removal of the endosperm and testaopposite the radicle tip of seeds resulted in root protrusion,the induction of free space and an increase of 4C DNA signalsin the three genotypes. It is concluded that ABA is crucialfor the efficient arrest of tomato embryo radicle tip cellsin G₁phase upon maturation, whereas GAs play an important rolein re-initiating 4C DNA levels upon germination. Dormancy; flow cytometry; free space; Lycopersicon esculentum ; maturation; priming; seed; tomato     

The Effects of Kinetin and Gibberellin on the Germination of Dehusked Seeds of Indica and Japonica Rice (Oryza sativaL.) under Anaerobic and Aerobic Conditions

The effects of kinetin and gibberellin were examined under anaerobicconditions (0% oxygen) and aerobic conditions (20% oxygen) onthe germination of dehusked seeds of indica and japonica ricecultivars that had been harvested at different times duringthe formation of seeds. Surjamkhi was used as a representativeof deep dormant indica cultivars and Assam IV as a less dormantindica cultivar. Sasanishiki was used as the japonica rice cultivar.Both phytohormones were applied at a concentration of 10^-3Mwhichproved to have the greatest stimulatory effect in preliminarywork at concentrations of 10^-3–10^-5M. Under aerobic conditions,inhibition of germination by dehusking of Sasanishiki seedsthat had been harvested either 30 or 60 d after anthesis wasovercome by kinetin and all seeds germinated. Complete germinationinduced by kinetin under aerobic conditions was also achievedwith the dehusked seeds of the indica rice cultivar Assam IVthat had been harvested on two occasions and of Surjamkhi thathad been harvested 28 d after anthesis. In contrast, germinationof dehusked japonica seeds stimulated by anaerobiosis was inhibitedby kinetin. The stimulatory effects of gibberellin on the germinationof indica and japonica rice seeds were observed under aerobicand anaerobic conditions. Under anaerobic conditions, the responsesof dehusked indica and japonica rice seeds to kinetin and gibberellindiffered, being negative with kinetin and positive with gibberellin.Under aerobic conditions, the stimulatory effects of kinetinon germination of dehusked seeds were greater than those ofgibberellin. Thus, treatment with kinetin appears to be usefulfor breaking the considerable dormancy commonly observed inthe dehusked seeds of indica rice. Mechanisms are proposed toexplain the stimulatory effects of these phytohormones on thegermination of dehusked seeds of indica and japonica rice underaerobic and anaerobic conditions. Rice; Oryza sativaL.; seed germination; dehusking treatment; gibberellin; indica; japonica; kinetin; oxygen; dormancy; germination inhibition; seed formation     

The Biosynthesis of Cytokinins in Germinating Lupin Seeds

Imbibed intact seeds, and excised embryos and cotyledons ofyellow lupin (Lupinus luteus L. cv. Weiko III) have been incubatedwith [¹⁴C]-adenine to investigate cytokinin biosynthesis duringthe early stages of germination. Following incubation the tissueswere extracted and purified by solvent partition and chromatographyon cellulose phosphate, diethylaminoethyl cellulose and SephadexLH-20 columns. Using a variety of thin layer chromatographic,high performance liquid chromato-graphic and chemical procedures,incorporation of ¹⁴C into dihydrozeatin riboside and its nucleotidewas demonstrated in extracts of intact embryos, intact cotyledonsand excised embryos. However, radioactivity was not found associatedwith cytokinins in fractions derived from the isolated cotyledons.This is the first direct demonstration of cytokinin biosynthesisin germinating seeds and the results indicate that the capacityfor cytokinin biosynthesis is probably confined to the embryonicaxes. If this is so, the levels of [¹⁴CJ-dihydrozeatin ribosideassociated with intact embryo and intact cotyledon fractionsindicate that the synthesized cytokinin is transported to andaccumulates in the cotyledons. Key words: Lupinus luteus, cytokinin biosynthesis, seed germination     

ACTION OF KINETIN ON PHOTOSENSITIVE GERMINATION OF LETTUCE SEED AS COMPARED WITH THAT OF GIBBERELLIC ACID

1. Experiments with the seeds of Grand Rapids lettuce showedthat the germination induced by gibberellic acid or by red lightis strongly accelerated by kinetin, although the latter itselfcan promote the germination in the dark only slightly. The reversiblelight reactions of the phytochrome system interact with kinetintreatment just as effectively as with water-imbibed controls.The site of primary action of red light is not altered by kinetintreatment. Kinetin does not modify the water uptake of the seedsfor at least 8 hours. 2. Kinetin was found to inhibit the growth of the hypocotyland root of the seed, but to promote very markedly the expansionof the cotyledons. This effect was observed not only with cotyledonsin intact seeds but also with isolated cotyledons. The expansionof kinetin-treated cotyledons is further promoted by red light,but not by far-red, as is also the case with germination itself. 3. A number of purine derivatives which have been reported topromote germination also cause expansion of isolated cotyledons. 4. Gibberellic acid promotes both hypocotyl elongation and cotyledonexpansion in the dark, but this effect does not interact withthe phytochrome system. The site of action of gibberellic acidprobably lies in the axis. 5. It is concluded, therefore, that the site of kinetin actionis in the cotyledons, whose expansion helps to break the seed-coatwhen light or gibberellin has contributed the primary stimulus. ¹ Present address: Johnson Foundation for Medical Physics, Universityof Pennsylvania, Philadelphia, Pa. (Received January 16, 1963; )     

多年生黑麦草种子萌发及植株再生的研究

对不同预处理条件下多年生黑麦草种子萌发、愈伤组织诱导以及植株再生进行了研究。结果表明,35℃恒温干燥处理3 h、浸泡16 h后,室温晾48 h的干湿交替及强度为1 000 GS磁场处理30 min均能使种子的发芽率得到显著提高。最适诱导培养基为MS+2,4-D 6 mg·L^-1,有机添加物对于愈伤组织的生长影响并不显著,但可以改善其状态;较为适宜的分化培养基为MS+2 mg·L^-1 6-BA或KT;生根培养基为附加0.5 mg·L^-1 NAA的MS培养基。     

Untersuchungen über die Regulation der DNS-Synthese während des Aktivitätswechsels vonAgrostemma githago-Samen

The relationships between DNA synthesis and germination capacity ofAgrostemma seeds have been studied. Protein synthesis and RNA synthesis are activated at the very beginning of imbibition, whereas DNA synthesis starts in the second part of the imbibition phase. Agrostemma seeds inhibited by higher temperature (30° C), or aged seeds with a low germination capacity are characterized by a remarkably reduced protein synthesis. DNA synthesis is also reduced. The inhibition of protein-synthesis ofAgrostemma embryos fed with cycloheximid or actinomycin D causes a depression of DNA synthesis. These results indicate that the initiation of DNA synthesis of imbibingAgrostemma seeds depends on the synthesis of special proteins. Abscisic acid inhibits growth as well as DNA synthesis of isolatedAgrostemma embryos. Mitomycin inhibits germination and DNA synthesis to the same extent. Dormant seeds with an undiminished intensity of protein synthesis also show a reduced incorporation of³H-thymidine in DNA. We suggest that DNA synthesis of imbibed seeds, which is a necessary prerequisite for the radicle protrusion, is involved in the mechanism of afterripening of theAgrostemma seeds.